Fig. 7. The effect of Ned1-overproduction on the nuclear structure. (A) A 3D image of a cell with overproduced Ned1. Electron micrographs from serial thin sections were used to construct the image (see Materials and Methods). The cell was incubated at 30°C for 24 hours to induce the expression of Ned1 protein. Blue, microtubule bundle in the nucleus; red, the spindle pole body (SPB). (B) Representing parts of electron micrographs used for construction of the 3D image shown in A. N, nucleus; MT, microtubules. (a) Middle part of the cell; arrowhead, the SPB situated in the cytoplasm. (b,c) Left and right parts of the cell, respectively; arrows indicate the ends of extended nuclear envelope. (C) Wild-type cells carrying both pREP82tubGFP and pREP1ned1⁺cDNA are incubated in the absence of thiamine at 30°C for 24 hours. (a,c) Localization of Nup189. (b,d) Microtubules (green), Nup189 (red) and DAPI (blue). (D) Suppression of the nuclear elongation by the pim1-46 mutation. pim1⁺ or pim1-46 cells were induced for full expression of Ned1 (26°C for 28.5 hours) and the nuclei were stained with DAPI. (E,F) Cells carrying either Cut12-GFP (E) or Mis6-GFP (F) were incubated as in A for the Ned1-overproduction (bottom panels; in upper panels cells without Ned1-overproduction) and stained with DAPI and anti-Sad1 antibody. Merged: cyan, DAPI; green, GFP; red, Sad1. Bars in B: 0.2 µm (a), 0.4 µm (b, c); other bars, 2 µm.

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